lacZ Antibody

SKU: orb344789

Description

Beta galactosidase antibody

Images & Validation

• Tested Applications: ELISA, IHC, WB
• Dilution Range: ELISA: 1:10,000, IHC: 1:1,500, WB: 1:5,000 - 1:10,000
• Application Notes:
  Anti-Beta-Gal Antibody has been tested by ELISA and western blot and is suitable for dot blot, immunofluorescence microscopy, immunoprecipitation, conjugation and most immunological methods requiring high titer and specificity. The antibody recognizes both frozen tissue sections, paraffin embedded tissue and 4% paraformaldehyde fixed tissue for most immunohistochemical analysis. A 1:1,500 dilution has been reported to detect beta-galactosidase in adult rat spinal cord tissue after infection with helper-dependent adenovirus expressing lacZ. In this particular experiment, tissue was perfused with 4% paraformaldehyde and cryostat-cut (35 µm) to produce free-floating sections.

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Immunogen: Full length native Beta Galactosidase isolated from E.coli
• Target: lacZ
• Purity: Beta-Galactosidase Antibody is an IgG fraction antibody purified from monospecific antiserum by a multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer stated above. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum as well as purified and partially purified Beta Galactosidase [E.coli]. Cross reactivity against Beta Galactosidase from other tissues and species may occur but have not been specifically determined. Very low background staining has been reported in various assays.
• Conjugation: Unconjugated

Storage & Handling

• Storage: Store vial at -20° C or below prior to opening. This vial contains a relatively low volume of reagent (25 µL). To minimize loss of volume dilute 1:10 by adding 225 µL of the buffer stated above directly to the vial. Recap, mix thoroughly and briefly centrifuge to collect the volume at the bottom of the vial. Use this intermediate dilution when calculating final dilutions as recommended below. Store the vial at -20°C or below after dilution. Avoid cycles of freezing and thawing.
• Form/Appearance: Liquid (sterile filtered)
• Buffer/Preservatives: Preservative: 0.01% (w/v) Sodium Azide. Stabilizer: None; Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Concentration: 1.0mg/mL
• Expiration Date: 12 months from date of receipt.
• Dry Ice Shipping: Please note: This product requires shipment on dry ice. A dry ice surcharge will apply.
• Disclaimer: For research use only

Alternative Names

rabbit anti-Beta Galactosidase Antibody, rabbit anti-beta gal antibody, ß-Gal, Anti-ß-Gal Antibody

Western Blot of Rabbit Anti-Beta-Galactosidase Antibody. Lane 1: Beta-Galactosidase Reduced [0.1 µg]. Lane 2: Opal Prestained Molecular Weight Marker. Lane 3: Beta-Galactosidase Non-Reduced [0.1 µg]. Primary Antibody: Anti-Beta-Galactosidase at 1:1000 overnight at 2-8°C. Secondary Antibody: Goat Anti-Rabbit IgG HRP (p/n orb347654) 1:70000 for 30 min at RT. Expected MW: ~117kDa.

Western Blot of Rabbit Anti-Beta-Galactosidase Antibody. Lane 1: partially purified preparation b-Galactosidase [1.0 µg]. Lane 2: Molecular Weight Marker. Primary Antibody: Anti-Beta-Galactosidase at 1:1000 overnight at 2-8°C. Secondary Antibody: Goat Anti-Rabbit IgG IRDye®800 1:10000 for 45min at RT. Observed MW: ~117kDa.

Western blotting using Biorbyt's Fluorescein conjugated anti-b-Galactosidase antibody shows a band at ~117 kDa (lanes 1 - 3) corresponding to 60 ng, 30 ng and 15 ng, respectively of b-Gal present in partially purified preparations (arrowhead). Lane 4 shows no cross reactivity with proteins present in a non-specific control E.coli lysate. Proteins were resolved on a 4-20% Tris-Glycine gel by SDS-PAGE and transferred to nitrocellulose and blocking using Blocking Buffer for Fluorescent Western Blotting (p/n orb348637). The membrane was probed with fluorescein conjugated anti-b-Galactosidase (p/n orb344848) diluted to 1:10000. Reaction occurred for 2 hours at room temperature. Molecular weight estimation was made by comparison to a prestained MW marker in lane M.

Quick Database Links

Gene Symbol

lacZ

UniProt

P00722

RefSeq (Protein):NP_414878.1